JE and Dengue .


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JE and Dengue . Panbio’s new JE-Dengue IgM Combo ELISA. Overview. Both Dengue and Japanese encephalitis (JE) belong to the Flavivirdae family and co-circulate in many parts of the world (Figure 1 – following slide ).
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Slide 1

JE and Dengue Panbio\'s new JE-Dengue IgM Combo ELISA

Slide 2

Overview Both Dengue and Japanese encephalitis (JE) have a place with the Flavivirdae family and co-course in many parts of the world (Figure 1 – taking after slide ). Serological refinement between these two flavivirus contaminations is frequently important however troublesome because of the level of cross-responsive epitopes inside the flavivirus family. Precise location of these infections is foremost to treatment and reconnaissance programs. Serological tests to help with the analysis of dengue infection contamination are generally accessible, however constrained business viral diagnostics for JE have been accessible.

Slide 3

#Adapted from maps created by the Centers for Disease Control World dispersion of Dengue – 2005 CDC http://www.cdc.gov/ncidod/dvbid/dengue/delineate 2005.htm Distribution of Japanese Encephalitis in Asia, 1970-1998 http://www.cdc.gov/ncidod/dvbid/jencephalitis/map.htm

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Japanese encephalitis 1 Japanese encephalitis (JE) infection: flavivirus antigenically identified with St. Louis encephalitis infection Transmitted through mosquito es of the Culex tritaeniorhynchus gathering Can bring about an adorable encephalitis which may advance to loss of motion, seizures, unconsciousness and passing , however t he dominant part of contaminations are subclinical Case-casualty proportion: 30% Serious neurologic sequela: 30% Leading reason for viral encephalitis in Asia with 30-50,000 cases announced every year Those at hazard include: Residents of rustic territories in endemic areas Active obligation military sent to endemic regions Expatriates in country regions Disease chance greatly low in voyagers

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Dengue 2 Dengue infection serotypes 1 – 4: flavivirus Transmission by means of mosquito ( Aedes aegypti ) Symptoms incorporate sudden onset of fever, extreme migraine, myalgias and arthralgias, leukopenia, thrombocytopenia and hemorrhagic signs Occasionally delivers stun and drain, prompting to death Incidence is variable relying upon pandemic movement Globally, there are an expected 50 to 100 million instances of dengue fever (DF) and a few hundred thousand instances of dengue hemorrhagic fever (DHF) every year Average case casualty rate of DHF is around 5% The illness is encountering a resurgence in the tropics and plagues are bigger and more successive

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Dengue and JE: the entangled analysis Early in disease dengue and JE can show likewise. fever, cerebral pain, sickness, spewing, and myalgia 4,5 Both are connected with patients having a background marked by mosquito presentation in endemic regions. Level of cross-reactivity among the Flavivirus amass has customarily entangled serological finding.

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Why recognize JE & dengue? Remedy finding is essential for general wellbeing observation Enable the exact appraisal of the JE load Ensure rectify treatment Although the treatment for both diseases is strong and in no hostile to viral treatment accessible, patients with JE may require encouraging, aviation route administration and seizure control 4 .

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JE-Dengue IgM Combo ELISA Intended utilize The Panbio Japanese Encephalitis - Dengue IgM Combo ELISA is for the subjective hypothetical location of IgM antibodies to Japanese encephalitis and dengue infection in serum as a guide in the clinical research facility conclusion of Japanese encephalitis and dengue infection disease in patients with clinical side effects steady with encephalitis or dengue fever. This examine is a serological guide to finding of Japanese encephalitis or dengue disease and positive outcomes ought to be affirmed by PRNT or ebb and flow CDC rules.

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Product situating Laboratory showcase Offer to research facilities as of now utilizing Panbio Dengue items Laboratories may keep utilizing the Panbio Dengue ELISAs or rapids to separate amongst essential and auxiliary dengue disease JE-Dengue IgM Combo ELISA does not separate amongst essential and optional dengue contamination Offer to labs at present alluding JE testing Use the JE-Dengue IgM Combo ELISA for JE finding fundamentally Areas where dengue co-circles

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Specifications Name: J apanese Encephalitis - Dengue IgM Combo ELISA Cat No: E-JED01C Expiry: 1 2 months from date of make Storage: 2 – 8 °C Antigen: Recombinant Dengue 1 – 4 Recombinant JE Antigen prep: Dilute 1/250 before utilize (discard any left-over weakened antigen) Sample: Serum Assay time: 2 hr 10 min add up to hatching time Kit estimate: 48 tests (96 well plate give, 2-wells required per persistent) Procedure: Standard Panbio catch ELISA organize

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Procedure E ach test is tried in copy, with one well utilizing JE-MAb complex and the other well utilizing Dengue-MAb complex in the second step. Weaken both antigens 1/250 utilizing the Antigen Diluent before utilize . Staying unused concentrated antigen ought to be put away at 2-8 ºC. Proceed according to the accompanying stream outline

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JE-Dengue IgM Combo ELISA S erum antibodies join with the counter human IgM covered on the plate. At the same time the peroxidase conjugated MAb and antigen provided shape buildings when hatched together. Washing expels remaining serum The antigen-Mab edifices tie if antigen-particular antibodies have been caught. The dreary substrate, tetramethylbenzidine/hydrogen peroxide (TMB/H2O2) is hydrolysed to a blue chromogen Stopping the hydrolytic response with corrosive turns the TMB yellow Color improvement demonstrates the nearness of particular antibodies in the test

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Interpretation # * Results ought to be affirmed by PRNT or current CDC rules. # Refer to Instructions for use for full elucidation rules

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Interpretation cont. On the off chance that positive on the dengue pack, this demonstrates a dengue infection contamination, regardless of the possibility that likewise positive on the JE unit. On the Panbio JE-Dengue IgM Combo ELISA cross-reactivity was overwhelmingly just discovered one way i.e. at the JE level. This cross-reactivity is a typical element of flavivirus immunizer tests and is normally more sensational when local infection arrangements are utilized. The particular recombinant infection antigens utilized as a part of this examine give unmistakable favorable circumstances over normal local infection arrangements particularly the dengue infection antigens. In this manner the two-well approach is basic to recognize dengue and JE.

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Performance: Study locales Reference research facility, Bangkok, Thailand Data utilized for bundle embed Inhouse examine Presented at JE Bi-Regional Meeting, B angkok,Thailand April 2005 AFRIMS, Thailand WHO Collaborating Center Competitive assessment Panbio versus 2 economically accessible ELISAs and an in-house ELISA

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Performance Evaluation by a reference research facility in Bangkok, Thailand Performance of the item was surveyed utilizing 360 sera gathered for testing at a reference lab in Bangkok, Thailand, and described utilizing the reference lab\'s in-house IgM and IgG ELISAs. 121 specimens JE positive 111 examples essential or auxiliary dengue tests 128 endemic specimens seronegative for both JE and dengue

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Good refinement amongst dengue and JE Only 2 dengue tests erroneously analyzed as JE Performance cont. Assessment by a reference research facility in Bangkok, Thailand

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Performance cont. Assessment by a reference research center in Bangkok, Thailand High affectability and specificity

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Performance Inhouse think about exhibited at JE Bi-Regional Meeting, Bangkok,Thailand April 2005 A populace of 349 portrayed examples was utilized to assess the execution of the JE-Dengue IgM Combo ELISA. USA and Australian ordinary contributor sera (n=265) Dengue IgM positive sera (n=19) JE IgM positive sera (n=65).

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Clear qualification amongst dengue and JE

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Performance cont. Inhouse think about displayed at JE Bi-Regional Meeting, Bangkok,Thailand April 2005

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Conclusions Inhouse ponder introduced at JE Bi-Regional Meeting, Bangkok,Thailand April 2005 The advancement of a 2-well ELISA using particular recombinant antigens gives a critical change in the determination of Japanese encephalitis. Test has an abnormal state of dengue and JE affectability and additionally specificity. In view of the abnormal state of specificity of the dengue recombinant antigen, a basic positive/negative correlation of the 2-well outcome can be utilized to decide a possible analysis. The configuration of the tests permits utilization of regular reagents with both JE and dengue antigens, including positive and negative control sera, upgrading convenience.

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Performance cont. USAMC-AFRIMS focused evaluation* April – July 2005 *Study has not yet been distributed

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Conclusions USAMC-AFRIMS aggressive assessment April – July 2005 Smaller number of steps included Performed superior to contenders High affectability & specificity High concurrence with very much perceived and set up in-house demonstrative pack Only monetarily accessible unit that separates amongst JE and dengue Panbio pack is profitable in territories where dengue co-circles

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Advantages Simple, institutionalized methodology same organization other Panbio catch ELISAs Short test time (2 hr 10 min) Good affectability and specificity inhouse and autonomous appraisal Excellent concurrence with flow indicative strategies Two-well approach permits serological separation amongst dengue and JE contamination Two ailments with one pack Recombinant antigens All 4 dengue serotypes are incorporated From Panbio: No. 1 in dengue analysis

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Promotional material Brochure – The way to separating amongst dengue and JE Flyer – ( PB0110 )

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Promotional material Poster exhibited at JE Bi-Regional Meeting, Bangkok,Thailand April 2005

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Competitors XCyton Diagnostics - JEV CheX pack 96 well ELISA Currently working just in specific parts of India Marketed by Qualigens Diagnos

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